CB2, PPAR-γ y GPR55 como dianas farmacológicas para un tratamiento anti-inflamatorio y neuroprotector en la enfermedad de Parkinson

Tesis inédita de la Universidad Complutense de Madrid, Facultad de Medicina, leída el 15-07-2022Parkinson’s disease (PD) is a chronic neurodegenerative disorder which courses with hypokinetic symptoms due to the selective death of the dopaminergic neurons of the substantia nigra pars compacta (SNpc) and the consequent dopaminergic denervation of the striatum. This disease is also a proteinopathy, as one of its features is the presence of aggregates of misfolded proteins, mainly α-synuclein, known as Lewy bodies (LB). The etiology of PD remains unknown, but several environmental and genetic risk factors have been described, together with aging as the main one. The main symptoms of PD are bradykinesia, rigidity, and resting tremor, usually determinant for the diagnosis, which appear when more than 50% of the dopaminergic neurons are already dead. However, there are non-motor symptoms such as sleep disturbances, mood symptoms, gastrointestinal problems, and olfactory dysfunction, which may be prodromal and could be useful for an early diagnosis. The pathological events that contribute to the neuronal death include not only protein aggregation, but also neuroinflammation, mitochondrial dysfunction, oxidative stress, and excitotoxicity. These mechanisms are known to be interconnected, worsening the dopaminergic neurodegeneration which disturbs the basal ganglia circuitry, preventing a correct motor functioning. The most frequent treatment is the replacement of dopamine levels, mainly with levodopa (L-DOPA), which recovers the mobility of the patients in the short term but causes irreversible dyskinesias after several years. This makes urgent the need of a disease-modifying treatment, and given the multifactorial pathophysiology of PD, we propose the use of pleiotropic molecules which can be effective at several levels, such as cannabinoids...La enfermedad de Parkinson (EP) es una enfermedad neurodegenerativa crónica que cursa con pérdida de movilidad por la muerte selectiva de las neuronas dopaminérgicas de la sustancia nigra pars compacta (SNpc) y la consiguiente denervación dopaminérgica del cuerpo estriado. Esta enfermedad es también una proteinopatía, ya que presenta agregados de proteínas mal plegadas, principalmente α-sinucleína, conocidos como cuerpos de Lewy (LB). La etiología de la EP sigue siendo desconocida, pero se han descrito varios factores de riesgo ambientales y genéticos, además del envejecimiento. Los principales síntomas de la EP son la bradiquinesia, la rigidez y el temblor en reposo, que son determinantes para el diagnóstico, y aparecen cuando más del 50% de las neuronas dopaminérgicas ya están muertas. Sin embargo, hay síntomas no motores, como alteraciones del sueño, trastornos del ánimo, problemas gastrointestinales y disfunción olfativa, que pueden ser prodrómicas y útiles para un diagnóstico precoz. Los eventos patológicos que contribuyen a la muerte neuronal incluyen la agregación proteica, pero también la neuroinflamación, la disfunción mitocondrial, el estrés oxidativo y la excitotoxicidad. Estos mecanismos están interconectados, agravando la neurodegeneración dopaminérgica que altera el circuito de los ganglios basales, impidiendo un correcto funcionamiento motor. El tratamiento más frecuente es la reposición de los niveles de dopamina con levodopa, que recupera la movilidad de los pacientes a corto plazo, pero provoca disquinesias irreversibles tras varios años. Esto hace necesario un tratamiento modificador de la enfermedad, y dada la fisiopatología multifactorial de la EP, se propone el uso de moléculas pleiotrópicas, como los cannabinoides...Fac. de MedicinaTRUEunpu

Beneficial effects of the phytocannabinoid Δ9-THCV in L-DOPA-induced dyskinesia in Parkinson's disease

The antioxidant and CB2 receptor agonist properties of Δ9-tetrahydrocannabivarin (Δ9-THCV) afforded neuroprotection in experimental Parkinson's disease (PD), whereas its CB1 receptor antagonist profile at doses lower than 5 mg/kg caused anti-hypokinetic effects. In the present study, we investigated the anti-dyskinetic potential of Δ9-THCV (administered i.p. at 2 mg/kg for two weeks), which had not been investigated before. This objective was investigated after inducing dyskinesia by repeated administration of L-DOPA (i.p. at 10 mg/kg) in a genetic model of dopaminergic deficiency, Pitx3ak mutant mice, which serves as a useful model for testing anti-dyskinetic agents. The daily treatment of these mice with L-DOPA for two weeks progressively increased the time spent in abnormal involuntary movements (AIMs) and elevated their horizontal and vertical activities (as measured in a computer-aided actimeter), signs that reflected the dyskinetic state of these mice. Interestingly, when combined with L-DOPA from the first injection, Δ9-THCV delayed the appearance of all these signs and decreased their intensity, with a reduction in the levels of FosB protein and the histone pAcH3 (measured by immunohistochemistry), which had previously been found to be elevated in the basal ganglia in L-DOPA-induced dyskinesia. In addition to the anti-dyskinetic effects of Δ9-THCV when administered at the onset of L-DOPA treatment, Δ9-THCV was also effective in attenuating the intensity of dyskinesia when administered for three consecutive days once these signs were already present (two weeks after the onset of L-DOPA treatment). In summary, our data support the anti-dyskinetic potential of Δ9-THCV, both to delay the occurrence and to attenuate the magnitude of dyskinetic signs. Although further studies are clearly required to determine the clinical significance of these data in humans, the results nevertheless situate Δ9-THCV in a promising position for developing a cannabinoid-based therapy for patients with PD.Fil: Espadas, Isabel. INSTITUTO CAJAL ; CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS;Fil: Keifman, Ettel. INSTITUTO CAJAL ; CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS; . Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Palomo Garo, Cristina. Universidad Complutense de Madrid; EspañaFil: Burgaz, Sonia. Universidad Complutense de Madrid; EspañaFil: García, Concepción. Universidad Complutense de Madrid; EspañaFil: Fernández Ruiz, Javier. Universidad Complutense de Madrid; EspañaFil: Moratalla, Rosario. INSTITUTO CAJAL ; CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS

Lichens and lichenicolous fungi of Serranía de Ronda (Málaga-Cádiz),southern Spain

As a result of a field trip organised by the Spanish Lichen Society in Serranía de Ronda, south Spain, a catalogue of 360 taxa is presented (336 lichens, 24 lichenicolous fungi). The list includes three new records for the Iberian Peninsula: Arthonia paretinaria, Micarea myriocarpa and Niesslia keissleri, 51new ones for the Autonomous Andalusian Community, and three and 81 new ones for the province of Cádiz and of Málaga, respectively. After these results, the total updated number of the province of Málaga rises to 556 lichens and lichenicolous fungi. The best represented lichen genus is Cladonia (18) with the most species, unlike Lecanora (15), Pertusaria (12), Physconia (12) and Collema (9). As regard habitat, most lichen species are mainly corticolous (55%), as opposed to saxicolous (24%), terricolous (14%) as the species growing on other lichens as lichenicolous fungi (7%). The percentages of lichen growth forms are mainly foliose (50%) and crustose (31%), while fruticose (7%), crustose squamulose (6%) and dimorphic (6%) are less represented. The lichen with a green photobiont (Chlorophyta 84%) predominates, while the cyanobacteria photobiont (15%) is less represented

Hongos liquenizados y liquenícolas de la Sierra de Albarracín (Teruel, España)

Se presenta un catálogo de 462 hongos liquenizados y liquenícolas de la Sierra de Albarracín (Teruel, Aragón, España), como resultado de la IV Campaña de Recolección organizada por la Sociedad Española de Liquenología (SEL). Diplotomma hedinii es novedad para la Península Ibé- rica y Lepraria leuckertiana constituye una segunda cita peninsular, 69 taxones son novedad para Aragón y 86 lo son para la provincia de Teruel

Lichens and lichenicolous fungi of Serranía de Ronda (Málaga-Cádiz), southern Spain

As a result of a field trip organised by the Spanish Lichen Society in Serranía de Ronda, south Spain, a catalogue of 360 taxa is presented (336 lichens, 24 lichenicolous fungi). The list includes three new records for the Iberian Peninsula: Arthonia paretinaria, Micarea myriocarpa and Niesslia keissleri, 51new ones for the Autonomous Andalusian Community, and three and 81 new ones for the province of Cádiz and of Málaga, respectively. After these results, the total updated number of the province of Málaga rises to 556 lichens and lichenicolous fungi. The best represented lichen genus is Cladonia (18) with the most species, unlike Lecanora (15), Pertusaria (12), Physconia (12) and Collema (9). As regard habitat, most lichen species are mainly corticolous (55%), as opposed to saxicolous (24%), terricolous (14%) as the species growing on other lichens as lichenicolous fungi (7%). The percentages of lichen growth forms are mainly foliose (50%) and crustose (31%), while fruticose (7%), crustose squamulose (6%) and dimorphic (6%) are less represented. The lichen with a green photobiont (Chlorophyta 84%) predominates, while the cyanobacteria photobiont (15%) is less represented

Lichenized and lichenicolous fungi from the Sierra de Albarracín (Teruel, Spain)

Se presenta un catálogo de 462 hongos liquenizados y liquenícolas de la Sierra de Albarracín (Teruel, Aragón, España), como resultado de la IV Campaña de Recolección organizada por la Sociedad Española de Liquenología (SEL). Diplotomma hedinii es novedad para la Península Ibérica y Lepraria leuckertiana constituye una segunda cita peninsular, 69 taxones son novedad para Aragón y 86 lo son para la provincia de Teruel.A catalogue of 462 lichenized and lichenicolous fungi from Sierra de Albarracín, (Teruel, Spain), is presented as the result of the SEL (Sociedad Española de Liquenología) IV collecting field trip. Diplotomma hedinii is recorded for the first time for the Iberian Peninsula and Lepraria leuckertiana is registered for the second time. The checklist also includes 69 taxa new for Aragón Community and 86 new for the province of Teruel

BiP Heterozigosity Aggravates Pathological Deterioration in Experimental Amyotrophic Lateral Sclerosis

In the present study, we investigated the involvement of the chaperone protein BiP (also known as GRP78 or Hspa5), a master regulator of intracellular proteostasis, in two mouse models of neurodegenerative diseases: amyotrophic lateral sclerosis (ALS) and Parkinson’s disease (PD). To this end, we used mice bearing partial genetic deletion of the BiP gene (BiP+/− mice), which, for the ALS model, were crossed with mutant SOD1 (mSOD1) transgenic mice to generate mSOD1/BiP+/− double mutant mice. Our data revealed a more intense neurological decline in the double mutants, reflected in a greater deterioration of the neurological score and rotarod performance, with also a reduced animal survival, compared to mSOD1 transgenic mice. Such worsening was associated with higher microglial (labelled with Iba-1 immunostaining) and, to a lesser extent, astroglial (labelled with GFAP immunostaining) immunoreactivities found in the double mutants, but not with a higher loss of spinal motor neurons (labelled with Nissl staining) in the spinal cord. The morphological analysis of Iba-1 and GFAP-positive cells revealed a higher presence of activated cells, characterized by elevated cell body size and shorter processes, in double mutants compared to mSOD1 mice with normal BiP expression. In the case of the PD model, BiP+/− mice were unilaterally lesioned with the parkinsonian neurotoxin 6-hydroxydopamine (6-OHDA). In this case, however, we did not detect a greater susceptibility to damage in mutant mice, as the motor defects caused by 6-OHDA in the pole test and the cylinder rearing test, as well as the losses in tyrosine hydroxylase-containing neurons and the elevated glial reactivity (labelled with CD68 and GFAP immunostaining) detected in the substantia nigra were of similar magnitude in BiP+/− mice compared with wildtype animals. Therefore, our findings support the view that a dysregulation of the protein BiP may contribute to ALS pathogenesis. As BiP has been recently related to cannabinoid type-1 (CB1) receptor function, our work also opens the door to future studies on a possible link between BiP and the neuroprotective effects of cannabinoids that have been widely reported in this neuropathological context. In support of this possibility, preliminary data indicate that CB1 receptor levels are significantly reduced in mSOD1 mice having partial deletion of BiP gene

Preclinical Investigation in Neuroprotective Effects of the GPR55 Ligand VCE-006.1 in Experimental Models of Parkinson’s Disease and Amyotrophic Lateral Sclerosis

Cannabinoids act as pleiotropic compounds exerting, among others, a broad-spectrum of neuroprotective effects. These effects have been investigated in the last years in different preclinical models of neurodegeneration, with the cannabinoid type-1 (CB1) and type-2 (CB2) receptors concen- trating an important part of this research. However, the issue has also been extended to additional targets that are also active for cannabinoids, such as the orphan G-protein receptor 55 (GPR55). In the present study, we investigated the neuroprotective potential of VCE-006.1, a chromenopyrazole derivative with biased orthosteric and positive allosteric modulator activity at GPR55, in murine models of two neurodegenerative diseases. First, we proved that VCE-006.1 alone could induce ERK1/2 activation and calcium mobilization, as well as increase cAMP response but only in the presence of lysophosphatidyl inositol. Next, we investigated this compound administered chronically in two neurotoxin-based models of Parkinson’s disease (PD), as well as in some cell-based models. VCE-006.1 was active in reversing the motor defects caused by 6-hydroxydopamine (6-OHDA) in the pole and the cylinder rearing tests, as well as the losses in tyrosine hydroxylase-containing neurons and the elevated glial reactivity detected in the substantia nigra. Similar cytoprotective effects were found in vitro in SH-SY5Y cells exposed to 6-OHDA. We also investigated VCE-006.1 in LPS-lesioned mice with similar beneficial effects, except against glial reactivity and associated inflammatory events, which remained unaltered, a fact confirmed in BV2 cells treated with LPS and VCE-006.1. We also analyzed GPR55 in these in vivo models with no changes in its gene expression, although GPR55 was down-regulated in BV2 cells treated with LPS, which may explain the lack of efficacy of VCE-006.1 in such an assay. Furthermore, we investigated VCE-006.1 in two genetic models of amyotrophic lateral sclerosis (ALS), mutant SOD1, or TDP-43 transgenic mice. Neither the neurological decline nor the deteriorated rotarod performance were prevented with this compound, and the same happened with the elevated microglial and astroglial reactivities, albeit modest spinal motor neuron preservation was achieved in both models. We also analyzed GPR55 in these in vivo models and found no changes in both TDP-43 transgenic and mSOD1 mice. Therefore, our findings support the view that targeting the GPR55 may afford neuroprotection in experimental PD, but not n ALS, thus stressing the specificities for the development of cannabinoid-based therapies in the different neurodegenerative disorders.This work has been supported by grants from CIBERNED (CB06/05/0089), MICIU (RTI- 2018-098885-B-100), ELA-Madrid-CM (B2017/BMD-3813), and Emerald Health Biotechnology-Spain. These agencies had no further role in study design, the collection, analysis, and interpretation of data, in the writing of the report, or in the decision to submit the paper for publicatioPeer reviewe

Hongos liquenizados y liquenícolas de la Sierra de Albarracín (Teruel, España)

[spa] Se presenta un catálogo de 462 hongos liquenizados y liquenícolas de la Sierra de Albarracín (Teruel, Aragón, España), como resultado de la IV Campaña de Recolección organizada por la Sociedad Española de Liquenología (SEL). Diplotomma hedinii es novedad para la Península Ibérica y Lepraria leuckertiana constituye una segunda cita peninsular, 69 taxones son novedad para Aragón y 86 lo son para la provincia de Teruel.[eng] A catalogue of 462 lichenized and lichenicolous fungi from Sierra de Albarracín, (Teruel, Spain), is presented as the result of the SEL (Sociedad Española de Liquenología) IV collecting field trip. Diplotomma hediniiis recorded for the first time for the Iberian Peninsula and Lepraria leuckertiana is registered for the second time. The checklist also includes 69 taxa new for Aragón Community and 86 new for the province of Teruel